Fusarium graminearum (teleomorph Gibberella zeae) is a significant pathogen of wheat and corn. F. graminearum forms multicellular macroconidia that play an important role in dissemination of the disease. The spatial pattern of morphogenesis in germinating macroconidia is described. Germ tubes preferentially emerge from the apical cells in a bipolar pattern that appears to be common to filamentous fungi. Chitin deposition occurs at two locations: the spore apices and cortical regions of macroconidial cells that subsequently produce a germ tube. The spatial pattern of morphogenesis requires the presence of functional microtubules, which may be responsible for the transport of key polarity factors to specific sites. These observations suggest that F. graminearum possesses a regulatory system that marks germ tube emergence sites. Perturbation of this system may represent an effective approach for inhibiting colonization of host plant surfaces.
Black yeasts are polyextremotolerant fungi that contain high amounts of melanin in their cell wall and maintain a primar yeast form. These fungi grow in xeric, nutrient depletes environments which implies that they require highly flexible metabolisms and have been suggested to contain the ability to form lichen-like mutualisms with nearby algae and bacteria. However, the exact ecological niche and interactions between these fungi and their surrounding community are not well understood. We have isolated 2 novel black yeasts from the genus Exophiala that were recovered from dryland biological soil crusts. Despite notable differences in colony and cellular morphology, both fungi appear to be members of the same species, which has been named Exophiala viscosa (i.e. E. viscosa JF 03-3 Goopy and E. viscosa JF 03-4F Slimy). A combination of whole genome sequencing, phenotypic experiments, and melanin regulation experiments have been performed on these isolates to fully characterize these fungi and help decipher their fundamental niche within the biological soil crust consortium. Our results reveal that E. viscosa is capable of utilizing a wide variety of carbon and nitrogen sources potentially derived from symbiotic microbes, can withstand many forms of abiotic stresses, and excretes melanin which can potentially provide ultraviolet resistance to the biological soil crust community. Besides the identification of a novel species within the genus Exophiala, our study also provides new insight into the regulation of melanin production in polyextremotolerant fungi.
Abstract One of the drawbacks during second-generation biofuel production from plant lignocellulosic biomass is the accumulation of glucose, the preferred carbon source of microorganisms, which causes the repression of hydrolytic enzyme secretion by industrially relevant filamentous fungi. Glucose sensing, subsequent transport and cellular signalling pathways have been barely elucidated in these organisms. This study therefore characterized the transcriptional response of the filamentous fungus Aspergillus nidulans to the presence of high and low glucose concentrations under continuous chemostat cultivation with the aim to identify novel factors involved in glucose sensing and signalling. Several transcription factor- and transporter-encoding genes were identified as being differentially regulated, including the previously characterized glucose and xylose transporter HxtB. HxtB was confirmed to be a low affinity glucose transporter, localizing to the plasma membrane under low- and high-glucose conditions. Furthermore, HxtB was shown to be involved in conidiation-related processes and may play a role in downstream glucose signalling. A gene predicted to encode the protein kinase PskA was also identified as being important for glucose metabolism. This study identified several proteins with predicted roles in glucose metabolic processes and provides a foundation for further investigation into the response of biotechnologically important filamentous fungi to glucose.
Summary The dimorphic fungus Candida albicans secretes farnesol, which acts as a quorum‐sensing molecule and prevents the yeast to mycelium conversion. In this study we examined the effect of farnesol in the filamentous fungus Aspergillus nidulans . We show that externally added farnesol has no effect on hyphal morphogenesis; instead, it triggers morphological features characteristic of apoptosis. Additional experiments suggest that mitochondria and reactive oxygen species (ROS) participate in farnesol‐induced apoptosis. Moreover, the effects of farnesol appear to be mediated by the FadA heterotrimeric G protein complex. Because A. nidulans does not secrete detectable amounts of farnesol, we propose that it responds to farnesol produced by other fungi. In agreement with this notion, growth and development were impaired in a farnesol‐dependent manner when A. nidulans was co‐cultivated with C. albicans . Taken together, our data suggest that farnesol, in addition to its quorum‐sensing function that regulates morphogenesis, is also employed by C. albicans To reduce competition from other microbes.
This chapter summarizes the progress achieved toward understanding the organization of fungal hyphae and the cellular systems involved in hyphal morphogenesis. Particular attention is paid to the mechanisms that have been implicated in the regulation of polarized growth and septum formation in filamentous fungi. Finally, the intriguing question of how morphogenetic regulatory systems may have evolved in the fungal kingdom is briefly addressed. Hyphal growth encompasses several different morphogenetic processes. Foremost among these is the establishment and maintenance of a stable axis of polarized growth. As a result, cell surface expansion and cell wall deposition are confined to a discrete location that ultimately becomes the hyphal tip. Genetic analyses demonstrate that Bni1 is absolutely essential for the establishment of hyphal polarity in A. gossypii. The importance of understanding the molecular mechanisms underlying polarized hyphal growth cannot be understated. The genetic tractability of filamentous fungi such as A. nidulans and N. crassa affords a tremendous opportunity to elucidate these mechanisms and to acquire insight that might be relevant to neurological disorders and other motor diseases. The use of increasingly sophisticated microscopy techniques has revealed the subcellular organization of hyphal tip cells and, in particular, emphasized the role of the Spitzenkörper in polarized hyphal growth. Future experiments that exploit genomic and proteomic tools will undoubtedly provide new insights that test the validity of this hypothesis and reveal the key symmetry-breaking event(s) that lead to polarized hyphal growth.
Tight control of the intracellular uracil level is believed to be important to reduce the occurrence of uracil incorporation into DNA. The pyrG gene of Aspergillus nidulans encodes orotidine 5'-phosphate decarboxylase, which catalyzes the conversion of orotidine monophosphate (OMP) to uridine monophosphate (UMP). In this study, we found that pyrG is critical for maintaining uracil at a low concentration in A. nidulans cells in the presence of exogenous uracil. Excess uracil and its derivatives had a stronger inhibitory effect on the growth of the pyrG89 mutant with defective OMP decarboxylase activity than on the growth of wild type, and induced sexual development in the pyrG89 mutant but not in wild type. Analysis of transcriptomic responses to excess uracil by digital gene expression profiling (DGE) revealed that genes related to sexual development were transcriptionally activated in the pyrG89 mutant but not in wild type. Quantitative analysis by HPLC showed that the cellular uracil level was 6.5 times higher in the pyrG89 mutant than in wild type in the presence of exogenous uracil. This study not only provides new information on uracil recycling and adaptation to excess uracil but also reveals the potential effects of OMP decarboxylase on fungal growth and development.
The importance of polarized growth for fungi has elicited significant effort directed at better understanding underlying mechanisms of polarization, with a focus on yeast systems. At sites of tip growth, multiple protein complexes assemble and coordinate to ensure that incoming building material reaches the appropriate destination sites, and polarized growth is maintained. One of these complexes is the polarisome that consists of Spa2, Bud6, Pea2, and Bni1 in Saccharomyces cerevisiae. Filamentous hyphae differ in their development and life style from yeasts and likely regulate polarized growth in a different way. This is expected to reflect on the composition and presence of protein complexes that assemble at the hyphal tip. In this study we searched for polarisome homologues in the model filamentous fungus Aspergillus nidulans and characterized the S. cerevisiae Spa2 and Bud6 homologues, SpaA and BudA. Compared to the S. cerevisiae Spa2, SpaA lacks domain II but has three additional domains that are conserved within filamentous fungi. Gene replacement strains and localization studies show that SpaA functions exclusively at the hyphal tip, while BudA functions at sites of septum formation and possibly at hyphal tips. We show that SpaA is not required for the assembly or maintenance of the Spitzenkörper. We propose that the core function of the polarisome in polarized growth is maintained but with different contributions of polarisome components to the process.